Elevation of oil levels in plants

a technology of oil content and plant, applied in the field of nucleic acid chemistry and agricultural biotechnology, can solve the problems of increasing the increasing the oil yield of transgenics, and not showing any evidence of increasing total fatty acid content, so as to increase the oil content, and improve the effect of oil production

Active Publication Date: 2005-01-06
MONSANTO TECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0012] In another embodiment, the present invention relates to a recombinant DNA construct, associated with increased oil production in plants, comprising a nucleic acid molecule encoding an HOI001 GBSS polypeptide operably linked to a promoter, which is functional in a plant cell.
[0013] The present invention describes and provides a method of increasing oil in a maize plant by expression of an HOI001 GBSS gene. This invention further provides a method of altering the kernel composition in a corn plant by expression of an HOI001 GBSS gene. This invention further describes and provides sequences of an HOI001 GBSS gene from Zea mays. This invention further provides vector constructs for plant transformation and tissue-specific expression of an HOI001 GBSS gene. This invention further provides maize plants transformed with the GBSS gene with higher oil levels when compared to plants with the same or similar genetic background, but not containing the inserted HOI001 GBSS gene. This invention further provides seeds from these maize plants. This invention further provides for kernels from maize plants transformed with the HOI001 GBSS gene containing a higher level of oil when compared to kernels from corn plants with the same or similar genetic background, but not containing the inserted HOI001 GBSS gene. This invention also provides oil and animal feed produced from these seeds and kernels.
[0014] The present invention further provides a method of marker-assisted breeding useful in breeding higher oil levels in maize.

Problems solved by technology

Although the use of these plant transgenes resulted in an increased production of lauric acid in canola and altered proportions of oleic acid in soy, there was no evidence of increased total fatty acid content, or increased oil yield in these transgenics.
However, the synthesis of fatty acids requires the coordinated activity of many enzymes, none of which when solely upregulated has been found to substantially increase oil content.

Method used

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  • Elevation of oil levels in plants
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Examples

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example 1

[0139] This example describes the isolation and sequencing of the HOI001 GBSS gene from corn line HOI001. HOI001 is an inbred plant derived from MGSC 915E (Maize Genetic Stock Center, Urbana, Ill.), and is more fully described in U.S. Patent Publication Nos. 20030172416 and 20030154524, both of which are incorporated herein by reference.

[0140] Genomic DNA was extracted from corn germ tissue from HOI001, 22 days after pollination, using the following procedure. Between 50-100 mg dissected germ tissue was placed in a Bio101 Multimix tube (Qiagen, Carlsbad, Calif., Cat. No. 657-601) with extraction buffer and glass beads. The extraction buffer consisted of 100 mM Tris-HCl (pH 8.0), 50 mM EDTA, 100 mM NaCl, 5 mM DTT, and 1% SDS. The tissue was then disrupted using the Bio 101 FASTPREP® machine (Qiagen) with 3 pulses of 20 seconds each. Following a 15 minute incubation at 65° C., 330 μl of 5M potassium acetate was added to each tube. The tubes were then incubated at 0° C. for 20 minutes...

example 2

[0207] This example sets forth the construction of plant transformation vectors containing the sequences of the HOI001 GBSS and the GBSS from inbred line LH59, [SEQ ID NOs: 1 and 7, respectively].

[0208] The HOI001 GBSS sequence was cut from the consensus-corrected version of pMON9480-2 using the restriction enzyme EcOR1. The resulting 4.7 kb fragment was purified following the manufacturer's protocol for the Qiagen miniprep kit (Qiagen, Inc., Valencia, Calif.). The ends of the fragment were blunted following manufacturer's protocol in the Stratagene PCR polishing kit (Stratagene, Inc., La Jolla, Calif.). The fragment was then gel purified using the Qiagen Gel Extraction kit (Qiagen), and cloned into pMON68203, a binary vector for plant transformation. The binary vector, pMON68203, contains left and right borders for T-DNA transfer, a CaMV 35S promoter::nptII::nos 3′ UTR plant selectable marker element (described in U.S. Pat. No. 6,255,560), and plant expression cassette sequences w...

example 3

[0210] This example describes the transformation of corn with the HOI001 GBSS and the GBSS from corn line LH59, using the vectors described in Example 2.

[0211] The transformation vectors pMON72506 and pMON72510 were used to transform maize plants using the following procedure.

[0212] Corn plants are grown in a greenhouse under standard practices. Controlled pollinations were made. The ears of the plants are harvested when the resulting hybrid embryos were 1.5 to 2.0 mm in length, usually 10-15 days after pollination. After removing the husks, the kernels on the ears were surface-sterilized by spraying with or soaking in 80% ethanol.

[0213] The Agrobacterium strain ABI, and an Agrobacterium tumefaciens binary vector system were used for the transformations. Plasmids pMON72506 and pMON72510 were transformed into Agrobacterium tumefaciens according to methods well known in the art. Prior to inoculation of corn cells the Agrobacterium cells are grown overnight at room temperature in AB...

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Abstract

This present invention provides a method for increasing oil levels in corn kernel tissue by expression of an HOI001 GBSS allele. The present invention also provides isolated nucleic acid molecules encoding a HOI001 GBSS polypeptide.

Description

[0001] This application claims the benefit of the filing date of the Provisional Application U.S. Ser. No. 60 / 483,491, filed Jun. 27, 2003, which is incorporated herein by reference.[0002] The present invention relates to the fields of nucleic acid chemistry and agricultural biotechnology. In particular, the present invention is directed at the identification of nucleic acids that encode proteins useful for increasing oil levels in maize plants and creating maize plants that include such nucleic acids. [0003] Plants are a major source of oils for feed, food, and industrial uses. While tissues of most plant species contain little oil, the cultivation of certain plant types, over many acres, permit large quantities of plant oils to be produced. If the oil content of these plants could be increased, then plant oils could be produced more efficiently. For example, the normal oil content of yellow #2, dent corn is about 4%. If the oil content of corn could be increased to 8% or even 12%,...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A23D9/00A23K1/14C12N9/10C12N15/82
CPCA23D9/00C12N15/8247C12N9/1051A23K1/14A23K10/30Y02A40/146
Inventor RAVANELLO, MONICAFOLEY, TERRYLEDEAUX, JOHNWYRICK, ANNETTESAVAGE, THOMAS
Owner MONSANTO TECH LLC
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